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目的:通过RNA干涉技术抑制肿瘤细胞端粒酶表达,探讨干涉后对肿瘤细胞生长的抑制作用.方法:根据人端粒酶逆转录酶(hTERT)mRNA编码序列,设计RNA干涉靶点,构建siRNA(smallinterferencingRNA)表达载体,并转染HeLa细胞,通过RT PCR法观察重组质粒转染肿瘤细胞后端粒酶mRNA、蛋白含量及细胞生长情况.结果:构建的siRNA表达载体可以使HeLa细胞端粒酶逆转录酶mRNA及其蛋白含量降低,转染质粒的细胞生长增殖速度减慢.结论:成功构建了针对人端粒酶逆转录酶的siRNA表达载体,通过转染HeLa细胞,可有效抑制细胞中端粒酶的表达,并引起细胞生长抑制.
OBJECTIVE: To inhibit the expression of telomerase in tumor cells by RNA interference and to investigate the inhibitory effect on the growth of tumor cells after intervention.Methods: According to the coding sequence of human telomerase reverse transcriptase (hTERT) mRNA, RNA interference target was designed and siRNA (smallinterferencingRNA) expression vector and transfected into HeLa cells.The expression of telomerase mRNA, protein and cell growth were detected by RT PCR.Results: The constructed siRNA expression vector can make the telomerase Reverse transcriptase mRNA and its protein content decreased, the transfected plasmids slowed down the growth of cell growth.Conclusion: The siRNA expression vector targeting human telomerase reverse transcriptase was successfully constructed and transfected into HeLa cells can effectively inhibit the proliferation of cells Telomerase expression and cell growth inhibition.